Kugu K, Ratts V S, Piquette G N, Tilly K I, Tao X J, Martimbeau S, Aberdeen G W, Krajewski S, Reed J C, Pepe G J, Albrecht E D, Tilly J L
Vincent Center for Reproductive Biology, Department of OB/GYN, Massachusetts General Hospital/Harvard Medical School, Boston, Massachusetts 02114, USA.
Cell Death Differ. 1998 Jan;5(1):67-76. doi: 10.1038/sj.cdd.4400316.
Recent data support a role for apoptosis, under tight regulatory control by bcl-2, oxidative stress response, tumor suppressor, and CASP gene family members, in mediating granulosa cell demise during follicular atresia in the rodent and avian ovary. Herein we evaluated the occurrence of apoptosis in the human and baboon ovary relative to follicular health status, and analyzed expression of several cell death genes in these tissues. In situlocalization of DNA strand breaks in fixed human and baboon ovarian tissue sections indicated that apoptosis was essentially restricted to granulosa cells of atretic antral follicles. Biochemical analysis of DNA oligonucleosomes in individual follicles isolated from baboon ovaries during the ovulatory phase revealed the presence of apoptotic DNA fragments in subordinate but not dominant follicles, thus substantiating the in situ labeling studies. Messenger RNA transcripts encoded by the bax death susceptibility gene, the bcl-xlong survival gene, the bcl-xshort pro-apoptosis gene, the p53 tumor suppressor gene, and two members of the CASP gene family (CASP-2/Ich-1, CASP-3/CPP32), were detected by Northern blot analysis of total RNA prepared either from human ovaries or from Percoll-purified granulosa-lutein cells obtained from patients undergoing assisted reproductive technologies. Lastly, immunohistochemical localization of the BAX death-susceptibility protein in the human ovary revealed abundant expression in granulosa cells of early atretic follicles, whereas BAX protein was extremely low or non-detectable in healthy or grossly-atretic follicles. We conclude that apoptosis occurs during, and is probably responsible for, folicular atresia in the human and baboon ovary. Moreover, apoptosis in the human ovary is likely controlled by altered expression of the same cohort of cell death regulatory factors recently implicated as primary determinants of apoptosis induction or suppression in the rodent ovary.
最近的数据支持凋亡在介导啮齿动物和鸟类卵巢卵泡闭锁过程中颗粒细胞死亡方面发挥作用，该过程受到bcl-2、氧化应激反应、肿瘤抑制因子和半胱天冬酶（CASP）基因家族成员的严格调控。在此，我们评估了人类和狒狒卵巢中凋亡的发生与卵泡健康状态的关系，并分析了这些组织中几种细胞死亡基因的表达。对固定的人类和狒狒卵巢组织切片中DNA链断裂的原位定位表明，凋亡基本上局限于闭锁窦卵泡的颗粒细胞。对排卵期从狒狒卵巢中分离出的单个卵泡进行DNA寡核小体的生化分析，结果显示在次级卵泡而非优势卵泡中存在凋亡DNA片段，从而证实了原位标记研究。通过对从人类卵巢或从接受辅助生殖技术患者获得的经Percoll纯化的颗粒黄体细胞制备的总RNA进行Northern印迹分析，检测到由bax死亡敏感性基因、bcl-x长生存基因、bcl-x短促凋亡基因、p53肿瘤抑制基因以及CASP基因家族的两个成员（CASP-2/Ich-1、CASP-3/CPP32）编码的信使RNA转录本。最后，人类卵巢中BAX死亡敏感性蛋白的免疫组织化学定位显示，在早期闭锁卵泡的颗粒细胞中大量表达，而在健康或严重闭锁卵泡中BAX蛋白极低或无法检测到。我们得出结论，凋亡发生在人类和狒狒卵巢的卵泡闭锁过程中，并且可能是其原因。此外，人类卵巢中的凋亡可能受一组细胞死亡调节因子表达改变的控制，这些因子最近被认为是啮齿动物卵巢中凋亡诱导或抑制的主要决定因素。
